Tag: pubmed

Zhongguo Zhong Yao Za Zhi. 2024 Oct;49(19):5288-5296. doi: 10.19540/j.cnki.cjcmm.20240712.702.

ABSTRACT

To investigate the effect of Buzhong Yiqi Decoction on regulatory T cells(Treg) in experimental rats with autoimmune thyroiditis(EAT) through the transforming growth factor-β(TGF-β)/Smad signaling pathway. Female SD rats were immunized with iodine-rich drinking water combined with Freund’s adjuvant and porcine thyroglobulin(pTG) to establish the EAT model of rats, and the levels of serum thyroperoxidase antibody(TPOAb) and thyroglobulin antibody(TGAb) were detected. Pathological sections by hematoxylin-eosin(HE) staining were observed. Treg in the rats’ spleen were extracted by immunomagnetic beads after the successful modeling and identified by flow cytometry. The extracted Treg were divided into blank group, Buzhong Yiqi Decoction group, TGF-β group, antagonist(LY3200882), and antagonist(LY3200882)+Buzhong Yiqi Decoction group. After the intervention, the cell counting kit-8(CCK-8) experiment was conducted to detect cell viability. Western blot and quantitative real-time PCR(RT-qPCR) were used to detect the expression of TGF-β/Smad signaling pathway-related proteins and genes. The results showed that the levels of TPOAb and TGAb increased in the rats in the model group compared to the rats in the blank group. HE staining showed that part of the follicles in the thyroid tissue of the rats in the model group were destroyed, and a large number of lymphocytes were infiltrated, indicating that the modeling was successful. After Treg were administered in vitro, CCK-8 results showed that the serum concentration of Buzhong Yiqi Decoction was below 40% to promote cell proliferation. The Buzhong Yiqi Decoction-containing serum group could increase the protein expression of TGF-β1, FoxP3, Smad2, and Smad4 compared with the blank serum group, while the expression of p-Smad2, p-Smad3, and Smad3 increased compared with the blank serum group, but the difference was not statistically significant. Compared with the antagonist group, the protein expressions of p-Smad2, Smad2, p-Smad3, Smad3, Smad4, and Smad7 did not significantly increase or decrease in the antagonist group after adding Buzhong Yiqi Decoction-containing serum. RT-qPCR showed that compared with the blank serum group, the mRNA expression of TGF-β1, FoxP3, Smad2, Smad3, Smad4, and Smad7 in the Buzhong Yiqi Decoction group increased or decreased in the same trend as that in the TGF-β group, but there was no statistical significance. After Buzhong Yiqi Decoction-containing serum was added to the antagonist group, the mRNA levels of TGF-β1, FoxP3, Smad2, Smad3, Smad4, and Smad7 were not statistically significant. In conclusion, Buzhong Yiqi Decoction could promote the stability and activity of Treg cells by promoting the secretion of TGF-β1 and regulating the expression of key signaling molecules TGF-β1, Smad2, and Smad4 in the TGF-β/Smad signaling pathway, thus affecting the immune balance of Th17/Treg and inhibiting the inflammatory response of rats with EAT.

PMID:39701767 | DOI:10.19540/j.cnki.cjcmm.20240712.702

Zhongguo Zhong Yao Za Zhi. 2024 Oct;49(19):5181-5192. doi: 10.19540/j.cnki.cjcmm.20240710.301.

ABSTRACT

This study aims to compare the effects and mechanisms of the standard decoction and formula granules of Paeoniae Radix Rubra in regulating the metabolism in the rat model of heat toxin and blood stasis. SD rats were randomized into control, model, standard decoction, and formula granules groups. After 14 days of administration, the rats in the latter three groups were subjected to subcutaneous injection with carrageenan and intraperitoneal injection with bacterial lipopolysaccharide for the modeling of heat toxin and blood stasis. The anal temperature, coagulation indexes, platelet aggregation(PAG), serum levels of thromboxane B_2(TXB_2), 6-ketone-prostaglondin F1α(6-keto-PGF1α), interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), von Willebrand factor(vWF), endothelial nitric oxide synthase(eNOS), and endothelin-1(ET-1) were measured. The metabolites in rat serum were identified by ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF-MS). Multivariate statistical analysis was performed to screen the differential metabolites, which were then subjected to the KEGG pathway enrichment analysis. The results showed that the standard decoction and formula granules of Paeoniae Radix Rubra improved the state of model rats by lowering the body temperature, inhibiting inflammation, reducing PAG, delaying coagulation, and regulating the vascular function. The metabolomics analysis screened out 15 and 65 differential metabolites in the standard decoction and formula granules groups, respectively, compared with the model group. The enrichment analysis of the differential metabolites showed that the standard decoction of Paeoniae Radix Rubra exerted effects by affecting four key metabolic pathways: α-linolenic acid metabolism, primary bile acid biosynthesis, linoleic acid metabolism, and phenylalanine metabolism. The formula granules of Paeoniae Radix Rubra exerted effects by affecting four key metabolic pathways: α-linolenic acid metabolism, citrate cycle(tricarboxylic acid cycle), purine metabolism, and glyoxylic acid and dicarboxylic acid metabolism. This study indicated that the standard decoction and formula granules of Paeoniae Radix Rubra had similar effects on the syndrome of heat toxin and blood stasis and both of them regulated α-linolenic acid catabolism, which provided a scientific basis for the clinical application of the formula granules of Paeoniae Radix Rubra.

PMID:39701756 | DOI:10.19540/j.cnki.cjcmm.20240710.301

Zhongguo Zhong Yao Za Zhi. 2024 Oct;49(20):5643-5651. doi: 10.19540/j.cnki.cjcmm.20240708.501.

ABSTRACT

This study aims to establish the implementation standard of health technology assessment(HTA) in traditional Chinese medicine(TCM), so as to promote the scientific and standardized implementation and application of HTA in TCM for high-quality decision-making. Based on the published HTA method guidelines, the literature analysis and expert consensus method were used to establish the implementation standard of HTA in TCM with implementation steps, assessment domains, and reporting checklists as the main content. A total of 41 HTA guidelines were included. After statistical induction and 4 consensus meetings, the preliminary standard of HTA in TCM was formed, and a complete TCM HTA implementation standard was established after two rounds of Delphi method, including 10 implementation steps(1 optional step and 9 necessary steps), 7 assessment domains(7 domains, 19 criteria and 37 sub-criteria, with 2 specific criteria and 5 specific sub-criteria for TCM diagnosis and treatment equipment or protocol health technologies), and 16 items in the reporting checklists. This study constructed a universal HTA implementation standard in TCM and incorporated patient perspectives. The implementation standard was normative, scientific, compatible, and considering the particularity of TCM. At the same time, the implementation standard was explained in detail, providing a reference for the practical application of HTA in TCM.

PMID:39701746 | DOI:10.19540/j.cnki.cjcmm.20240708.501

Zhongguo Zhong Yao Za Zhi. 2024 Oct;49(20):5451-5459. doi: 10.19540/j.cnki.cjcmm.20240717.301.

ABSTRACT

To analyze the differences and dynamic changes in the color, odor, and major chemical components of Astragali Radix during honey processing, this study used a CM-5 spectrophotometer and Hercules NEO ultra-fast gas phase electronic nose for analysis. High performance liquid chromatography was employed to determine the content of calycosin, calycosin-7-glucoside, formononetin, and ononin in the Astragali Radix decoction pieces processed with honey to different degrees. Multivariate statistical analyses including partial least squares-discriminant analysis(PLS-DA), orthogonal partial least squares-discriminant analysis(OPLS-DA), differential factor analysis(DFA), and Bayesian discriminant analysis were adopted to differentiate the Astragali Radix decoction pieces processed with honey to different degrees and evaluate the correlations between visual characteristics and chemical composition. The results showed that chromatic values c~*, b~*, and L~* were the main color difference variables distinguishing raw and honey-processed Astragali Radix decoction pieces. The reference ranges for the chromatic values of raw and honey-processed Astragali Radix decoction pieces were determined based on these key color parameters. A total of 18 odor components were identified in Astragali Radix during the honey-processing process, with cyclohexane and dimethyl sulfoxide identified as odor markers for distinguishing Astragali Radix decoction pieces with different honey-processing degrees. The content of calycosin had a significant correlation with that of 2-methylfuran and delta-nonalactone, two odor components. This study successfully and rapidly differentiated the Astragali Radix decoction pieces processed with honey to different degrees, providing a reference for the quality control and the quality evaluation of Astragali Radix decoction pieces based on the appearance traits during the process of honey processing.

PMID:39701728 | DOI:10.19540/j.cnki.cjcmm.20240717.301

Zhongguo Zhong Yao Za Zhi. 2024 Nov;49(22):6224-6234. doi: 10.19540/j.cnki.cjcmm.20240815.503.

ABSTRACT

This study systematically reviewed the randomized controlled trial(RCT) of traditional Chinese medicine(TCM) treatment of coronary heart disease patients with angina pectoris after percutaneous coronary intervention(PCI). The basic elements of these RCTs, including sample size and estimation method, randomizing scheme, allocation concealment, blind method implementation, data integrity, statistical method, TCM syndrome, intervention measures, treatment course, follow-up time, and outcome indicators, were analyzed to provide reference for the design of future RCT and the clinical application of TCM in treating angina pectoris after PCI. CNKI, Wanfang, VIP, SinoMed, PubMed, EMbase, Cochrane Library, Web of Science, ClinicalTrials.gov, and Chinese Clinical Trial Registry were searched for the RCT about TCM treatment of coronary heart disease patients with angina pectoris after PCI according to pre-defined criteria, with the time interval from inception to January 31, 2024. A total of 188 RCTs were included, of which 184 were clinical research articles and 4 were clinical trial registration schemes. These RCTs involved a total of 15 521 patients, with an average sample size of 83 patients and a maximum sample size of 248 patients. Among them, 126 RCTs reported TCM syndromes, the top three of which were Qi deficiency and blood stasis(38.89%), phlegm combined with stasis(17.46%), and Qi stagnation and blood stasis(9.52%). The control group received guideline-directed medical therapy(GDMT) or GDMT combined with placebo, and the treatment group received GDMT combined with TCM. The treatment mainly lasted for 4-8 weeks, most of the RCTs did not set the follow-up period or the follow-up period was unknown. A total of 160 outcome indicators were used, with the total frequency of 1 348. According to functional attributes, the outcome indicators can be categorized into 6 groups: symptoms/signs(403, 29.90%), TCM syndromes/symptoms(182, 13.50%), physical and chemical examination(468, 34.72%), quality of life(89, 6.60%), long-term prognosis(5, 0.37%), and safety evaluation(201, 14.91%). The clinical trial design of TCM intervention in angina pectoris after PCI of coronary heart disease is becoming more and more rigorous, while it remains to be improved. It is expected that more clinical trial schemes with rigorous design and taking into account the TCM advantages can be adopted in the future to provide a basis for the TCM treatment of angina pectoris after PCI of coronary heart disease.

PMID:39701718 | DOI:10.19540/j.cnki.cjcmm.20240815.503

Zhongguo Zhong Yao Za Zhi. 2024 Nov;49(22):6198-6213. doi: 10.19540/j.cnki.cjcmm.20240815.501.

ABSTRACT

Bayesian network Meta-analysis was conducted to assess the efficacy and safety of different Chinese medicine injections for dilated cardiomyopathy(DCM). CNKI, Wanfang, VIP, SinoMed, PubMed, Web of Science, EMbase, Cochrane Library, ProQuest, Cochrane Central Register of Controlled Trials, ClinicalTrials.gov were searched for the randomized controlled trial(RCT) from the inception to January 2024. The quality of the included RCT was evaluated using the Cochrane’s risk of bias assessment tool, and the quality of evidence for outcomes was assessed by GRADE. Stata 16.0 and RevMan 5.4 softwares were used for Meta-analysis. A total of 57 RCTs involving 4 812 subjects, 6 Chinese medicine injections and 6 outcome indicators were included. Bayesian network Meta-analysis indicated that:(1) In terms of increasing clinical total effectiveness rate, except Shenqi Fuzheng Injection + conventional western medicine, other 5 Chinese medicine injections combined with conventional western medicine outperformed conventional western medicine alone, and Xinmailong Injection + conventional western medicine had the best effect.(2) In terms of improving left ventricular ejection fraction, all of the 6 Chinese medicine injections combined with conventional western medicine were superior to conventional western medicine alone, and Shengmai Injection + conventional western medicine had the highest likelihood of being the best intervention.(3) On account of reducing left ventricular end-diastolic dimension, Shenfu Injection + conventional western medicine, Shenmai Injection + conventional western medicine, Huangqi Injection + conventional western medicine, and Xinmailong Injection + conventional western medicine outperformed conventional western medicine alone except Shengmai Injection + conventional western medicine, and Huangqi Injection + conventional western medicine had the highest probability of being the best treatment.(4) In terms of decreasing BNP, Huangqi Injection + conventional western medicine, Shenfu Injection + conventional western medicine, and Shenmai Injection + conventional western medicine were better than conventional western medicine alone, and Huangqi Injection + conventional western medicine had the highest likelihood of being the best treatment.(5) Considering the improvement in 6-min walk test, Shenmai Injection + conventional western medicine were superior to conventional western medicine alone.(6) In the case of improving cardiac output, Shenmai Injection + conventional western medicine, Shengmai Injection + conventional western medicine, Xinmailong Injection + conventional western medicine, and Shenqi Fuzheng Injection + conventional western medicine outperformed conventional western medicine alone except Shenfu Injection + conventional western medicine, thereinto, Shenmai Injection + conventional western medicine might be the optimal choice for increasing cardiac output level.(7) For safety, no statistically significant difference in adverse events was recorded between the experimental group and the control group(P=0.24). Bayesian network Meta-analysis showed that the addition of Chinese medicine injections to conventional western medicine exerted a more positive effect for DCM, and had advantages in alleviating clinical symptoms and improving cardiac function. However, due to the overall low quality of the included articles, the low inclusion of certain Chinese medicine injections, and the lack of direct comparison between different Chinese medicine injections, the results need to be further confirmed.

PMID:39701716 | DOI:10.19540/j.cnki.cjcmm.20240815.501

Zhongguo Zhong Yao Za Zhi. 2024 Nov;49(22):6138-6148. doi: 10.19540/j.cnki.cjcmm.20240809.301.

ABSTRACT

This study utilized ultra-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) to rapidly analyze and identify the chemical constituents in five processed products of Strychni Semen(raw, sand-roasted, fried, urine-soaked, and vinegar-processed products). Using PeakView software to extract compound information, 50 chemical components were identified based on retention time, accurate molecular ion peaks, secondary mass spectrometry data, and comparison with reference standards and relevant literature. Specifically, 41 components were identified in raw Strychni Semen, 48 in sand-roasted, 43 in fried, 41 in urine-soaked, and 40 in vinegar-processed products. Principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) were employed to identify differential components among the processed products, with variable importance in projection(VIP) values>1 and a t-test with P<0.05 as criteria. PCA revealed significant differences among the five processed products, demonstrating good clustering and separation. OPLS-DA identified 13 differential components, including brucine, strychnine, loganic acid, and chlorogenic acid. The results indicated that S. nux-vomica primarily contained alkaloids, organic acids, and glycosides, with significant compositional differences among the various processed products. Notably, the content of key components such as brucine and strychnine decreased after processing, with the most significant reduction observed in vinegar-processed Strychni Semen. In conclusion, this study provides a comprehensive comparison of the effects of different processing methods on the chemical composition of Strychni Semen, exploring the differences in the material basis of the processed products and offering data to support the scientific rationale behind the processing of Strychni Semen.

PMID:39701710 | DOI:10.19540/j.cnki.cjcmm.20240809.301

Zhongguo Zhong Yao Za Zhi. 2024 Sep;49(18):5045-5054. doi: 10.19540/j.cnki.cjcmm.20240604.501.

ABSTRACT

Based on the latent structure model and association rule analysis, the regularity of prescription for hyperlipidemia comorbid with hypertension was explored, and traditional Chinese medicine(TCM) pattern type was speculated to provide a reference for clinical pattern differentiation and treatment, and guideline establishment. The literature on the treatment of hyperlipidemia comorbid with hypertension by TCM was systematically searched on CNKI, Wanfang, VIP, and SinoMed academic platforms. The relevant information was extracted and a database was established using Microsoft Excel 2019. The frequency and properties of high-frequency TCM were statistically analyzed using Lantern 5.0 and RStudio software. The latent structure model and association rule analysis were applied to explore the medication regularity and speculate TCM pattern type. A total of 97 TCM prescriptions involving 104 TCM were included, with a cumulative frequency of 1 189 times. The high-frequency TCM included Poria, Gastrodiae Rhizoma, Atractylodis Macrocephalae Rhizoma, Pinelliae Rhizoma, Citri Reticulatae Pericarpium. The efficacy was mainly based on tonifying deficiency drugs, activating blood circulation drugs, and calming liver wind drugs. A total of 13 latent variables, 42 latent classes, and 5 comprehensive clustering models were obtained from the latent structure analysis. A total of 17 core formulas were obtained, suggesting that there were 5 common patterns in hyperlipidemia comorbid with hypertension, namely phlegm-dampness and blood stasis pattern, phlegm-blood stasis pattern, liver-fire hyperactivity pattern, Yin deficiency and Yang hyperactivity pattern, and liver and kidney deficiency pattern. Association rule analysis obtained 26 strong association rules, among which the highest support degree was Gastrodiae Rhizoma→Poria. Hyperlipidemia comorbid with hypertension belongs to the pattern of deficiency in the root and excess in the branch, which is closely related to the dysfunction of the liver, kidney, and spleen in the body. Phlegm-dampness and blood stasis, liver and kidney Yin deficiency are the main pathogenesis of this disease. The treatment should be based on the principles of calming liver fire, resolving phlegm, and strengthening spleen function. According to the pattern, Qi and blood as well as Yin and Yang should be coordinated to eliminate phlegm, blood stasis, etc., supplemented by treatments such as calming liver fire and eliminating wind from the liver.

PMID:39701687 | DOI:10.19540/j.cnki.cjcmm.20240604.501

Hum Reprod. 2024 Dec 19:deae269. doi: 10.1093/humrep/deae269. Online ahead of print.

ABSTRACT

STUDY QUESTION: Is there an association between the somatic loss of PTEN (phosphatase and tensin homolog) and ARID1A (AT-rich interaction domain 1A) and endometriosis disease severity and worse clinical outcomes?

SUMMARY ANSWER: Somatic PTEN loss in endometriosis epithelium was associated with greater disease burden and subsequent surgical complexity.

WHAT IS KNOWN ALREADY: Somatic cancer-driver mutations including those involving the PTEN and ARID1A genes exist in endometriosis without cancer; however, their clinical impact remains unclear.

STUDY DESIGN, SIZE, DURATION: This prospective longitudinal study involved endometriosis tissue and clinical data from 126 participants who underwent surgery at a tertiary center for endometriosis (2013-2017), with a follow-up period of 5-9 years.

PARTICIPANTS/MATERIALS, SETTING, METHODS: PTEN and ARID1A loss was assessed using established immunohistochemistry (IHC) methods as proxies for somatic loss by two independent raters. PTEN and ARID1A status for each participant was defined as loss (loss in at least one sample for a participant) or retained (no loss in all samples for a participant). Primary analyses examined associations between PTEN and ARID1A loss and disease burden based on anatomic subtype (superficial peritoneal endometriosis (SUP), deep endometriosis (DE), ovarian endometrioma (OMA)) and rASRM stage (I-IV). Secondary analyses explored associations of PTEN and ARID1A loss with demographics, surgical difficulty, and pain scores (baseline and follow-up). Additionally, using previously published data on KRAS codon 12 mutations for this cohort, we investigated associations between variables in the primary and secondary analyses and acquiring two or more somatic events (PTEN loss, ARID1A loss, or KRAS mutation) in this cohort. The risk of reoperation over the 5-9 years was also examined.

MAIN RESULTS AND THE ROLE OF CHANCE: PTEN loss (68.3%; 86 participants) exceeded ARID1A loss (24.6%; 31 participants). Inter-rater reliability was substantial for PTEN (k = 0.69; 95% CI: 0.62-0.77) and ARID1A (k = 0.64; 95% CI: 0.51-0.77). PTEN loss was significantly associated with more severe anatomic subtypes (P < 0.001; participants with SUP only = 46.4%; participants with DE only or OMA only = 72.7%; participants with mixed subtypes = 85.1%), and higher stages (P = 0.024; Stage I = 47.8%; Stage II = 73.7%; Stage III = 80.8%; Stage IV = 81.0%). Results were similar for ARID1A loss, albeit with smaller sample size limiting power. PTEN loss was further associated with non-White ethnicities (P = 0.017) and greater surgical difficulty (more frequent need for ureterolysis) (P = 0.02). There were no differences in pain scores (baseline or follow-up) based on PTEN or ARID1A status. Reoperation was uncommon (13.5% of the cohort), and patterns in reoperation rates based on the presence of somatic alterations did not reach statistical significance.

LIMITATIONS, REASONS FOR CAUTION: Sequencing was not performed to determine the type of PTEN and ARID1A somatic mutations resulting in loss of expression.

WIDER IMPLICATIONS OF THE FINDINGS: These results demonstrate a link between PTEN somatic loss and greater endometriosis disease burden. These findings underscore the potential relevance of PTEN loss and other somatic driver mutations in a future molecular classification of endometriosis.

STUDY FUNDING/COMPETING INTEREST(S): This study was funded by Canadian Institutes of Health Research (CIHR) project grant (MOP-142273 and PJT-156084). P.J.Y. was supported by a Health Professional Investigator award from Michael Smith Health Research BC, Canada, and a Canada Research Chair (Tier 2) in Endometriosis and Pelvic Pain. M.S.A. was supported by a Michael Smith Health Research BC Scholar award, and CIHR project grants (369990, 462997, and 456767). The sponsors did not play any role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; and in the decision to submit the article for publication. C.A. declares receiving payment from Pfizer for a symposium; being on advisory boards for AbbVie and Pfizer; being President and past President of the Canadian Society for the Advancement of Gynecologic Excellence (CanSAGE), co-lead of EndoAct Canada, and a board member of IPPS. M.A.B. has received consulting fees from AbbVie and Pfizer and grants from Ferring outside the scope of this work. D.G.H. is the founder of Canxeia Health but has no current affiliation. M.K. has received consulting fees from Helix Biopharma outside the scope of this work. M.S.A. received reimbursement of travel and registration fees to attend and present at the 2023 and 2024 annual meetings for the Society for Reproductive Investigation (SRI). P.J.Y. declares receiving: payment for a lecture from the International Society for the Study of Women’s Sexual Health (ISSWSH); honoraria from the CIHR; support to attend meetings from CanSAGE, ISSWSH, the International Pelvic Pain Society, the World Endometriosis Society (WES), the Society for the Study of Reproduction, and the Vulvodynia Summit; and discounted devices from Ohnut Wearable for a clinical trial. P.J.Y. is a data safety monitoring board member of a clinical trial funded by CIHR; and a strategic advisory board member for the Women’s Health Research Institute. P.J.Y. served as a board of directors member for CanSAGE and ISSWSH; was a junior board of directors member for WES; is a current board of directors member for WES; and was a committee chair for the Society of Obstetricians and Gynaecologists of Canada. A subset of these results was presented by the first author at the 71st Society for Reproductive Investigation Annual Scientific Meeting on 15 March 2024. Other authors have nothing to declare.

TRIAL REGISTRATION NUMBER: N/A.

PMID:39701665 | DOI:10.1093/humrep/deae269

Vet Comp Oncol. 2024 Dec 19. doi: 10.1111/vco.13036. Online ahead of print.

ABSTRACT

Mast cell tumours (MCTs) are the most frequent cutaneous neoplasia of the dog, and they have very variable biological behaviour and survival times. Surgery is still the best treatment, and despite the several adjuvant therapies described, many cases are very aggressive and resistant to these treatments making it urgent to find new therapeutic targets. Nowadays, immunotherapy targeting immune checkpoints has been described as a complementary treatment for several human cancers, but it is still very scarcely studied in veterinary medicine. Therefore, this study aimed to investigate the expression of the checkpoint proteins programmed death-ligand 1 (PD-L1) and cytotoxic T-lymphocyte antigen-4 (CTLA-4) to evaluate their potential as therapeutic targets for MCT. Through immunohistochemical study, it was analysed the expression of PD-L1 and CTLA-4 in 74 MCT cases from the archive of the Veterinary Pathology Laboratory of the University of Porto (LabPatVet). Tumour size, histological grade, ki-67 proliferation index, mitotic count and presence of metastatic disease were also assessed. Most of the cases expressed both immune checkpoints in neoplastic cells. There was a statistically significant inverse association between the expression of CTLA-4 and MCT grade (p < 0,001) and mitotic count (p < 0.001). PD-L1 was significantly and negatively related to HG (p = 0.004), and tumour size (р = 0.014). Tumour size, histological grade and mitotic count were positively associated with metastatic disease. Additionally, it was observed that the expression of PD-L1 and CTLA-4 was interrelated (p < 0.001). This study demonstrated that MCT cells express both PD-L1 and CTLA-4 and that their expression was associated with MCT prognostic factors.

PMID:39701664 | DOI:10.1111/vco.13036